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Analytical methods for qualitative and quantitative determination of PHA in cyanobacteria
Černayová, Diana ; Samek, Ota (referee) ; Sedláček, Petr (advisor)
The diploma thesis is confused to verify the applicability of selected physicochemical and spectroscopic methods for characterization of cyanobacteria, with special emphasis on possibilities of qualitative and quantitative analysis of polyhydroxyalkanoates (specifically polyhydroxybutyrate (PHB)) accumulated in cyanobacterial cells. The sample basis of the work was formed by cultures of cyanobacterial strains of Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. The cultures were were cultivated in several ways to cover the widest possible range of physiological conditions and PHB contents, in particular using an autotrophic way of cultivation on shakers and multicultural culture method in a basic culture medium,and in media enriched with 2% salt (NaCl ) as well as mixotrophic culture media with different types of the carbon substrate. After few weeks of cultivation, cyanobacterial cultures were obtained and complexly analyzed by following techniques- cell suspensions were analyzed by flow cytometry and UV-VIS spectrometry (transmission and diffusion transmission mode), dry cell biomass was characterised by gas chromatography to obtain a exact amount of PHB, and then FT-IR spectrometry and thermogravimetric analysis. The work aimed to assess whether any of these methods can be a quick and affordable alternative to the determination of PHB content to the most commonly used method of gas chromatography, but also to assess what additional information about the physiological state of cyanobacterial cells can provide test methods. The highest correlation on PHB content was determined for the parameters determined by infrared spectroscopy, in which specific peaks from the characteristic wavelengths for polyhydroxybutyrate were important. Weak correlations on PHB content were achieved in thermogravimetric analysis and cytometry, using the hydrophobic fluorescent probe BODIPY 439/503, which bound to lipophilic parts of cells. In addition to the determination of PHB, it was possible to determine pigments present in cyanobacteria (such as chlorophyll, phycocyanin and carotenoids) by flow cytometry and UV-VIS diffusion transmission spectrometry. In the end, results from all used techniques were compared by PCA analysis to determine the similarity of all analyzed samples.
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Thin films prepared in RF glow discharge and their physico-chemical properties
Bránecký, Martin ; Boušek, Jaroslav (referee) ; Čech, Vladimír (advisor)
Theoretical part of this master thesis was focused on literature recherché dealing with the formation of thin films, plasma, plasma analyses using mass spectrometry and plasma polymerization. Further, this section describes the analysis of thin films using optical methods such as spectroscopic ellipsometry and FT-IR spectrometry. Experimental part describes the materials which are used for the preparation of thin films as well as a description of the equipment for preparation of thin films using a technology of plasma-enhanced chemical vapor deposition (PE-CVD). Control of deposition conditions and monitoring of plasma with its products result in high reproducibility of thin films. The last part of the thesis describes the results of measurement of the first group of samples and their ellipsometric, mass spectrometry and FT-IR evaluation with respect to the deposition conditions.
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Phosphate binders
Kalina, Lukáš ; Ptáček, Petr (referee) ; Šoukal, František (advisor)
The work deals with the preparation and study of polyphosphate composites. Aluminum phosphates provide some unusual properties like high-heat resistance in terms of inorganic binders. The addition of chromium in the form of Cr2O3 or directly the bond with the aluminum-chromium-phosphate can improve the properties of this binder. Highly viscous Al2Cr(H2PO4)9 and Al3Cr(H2PO4)12 binders were prepared by dissolving Al(OH)3 and CrO3 in 85% phosphoric acid, and mixed with Al2O3 and Cr2O3 fillers. The composites were cured in the furnace at 150 °C for 24 h under pressure of 10 MPa. During the annealing at temperatures up to 1 500 °C changes in chemical structure of the cured composites were observed, leading to the understanding of the creation of crystalline phases and their eventual changes. The characterization of binders was mainly based on FT-IR, EDAX, TG-DTA and optical microscopy analyses.
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Testing of processing stability of PP-g-IA
Hampapa, Břetislav ; Tocháček, Jiří (referee) ; Kučera, František (advisor)
The Bachelor’s study is interested in processing stability of polypropylene grafting with Itaconic anhydride. A present knowledge about thermal degradation of Itaconic derivatives was summarized in theoretical part. Samples were prepared by multiple extrusions technique from virgin PP (Borsodchem, Hungary) and PP-g-IA. Die temperature 230 °C, processing time 4 minutes by 20 RPM. Qualitative and quantity analysis was made by FT-IR spectroscopy.
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Synthesis of SiO2/Au core/shell nanoparticles for enhancement of OCT signal
Kantorová, Martina ; Kolář, Radim (referee) ; Drbohlavová, Jana (advisor)
The aim of this master thesis is synthesis of SiO2/Au core/shell nanoparticles. These nanoparticles should improve the signal of optical coherence tomography (OCT). The procedure of nanoparticle synthesis is described in the work. Synthetized nanoparticles were analyzed by scanning electron microscopy (SEM), Ultraviolet-Visible spectroscopy (UV-Vis), Near-infrared spectroscopy (NIRS), Fourier transform-infrared spectroscopy (FT-IR) and Dynamic light scattering (DLS). Then, the synthetized nanoparticles were tested for enhancement of OCT signal. For this testing were also used commercial nanoparticles with the same structure and similar size.
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Properties of aminoclay complexes and biologically active substances
Dušek, Jakub ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This paper builds on previous research of aminoclay complexes in undergraduate studies. Theoretical part deals with study of current problems of aminoclay complexes with bioactive substances and the choice of substances for complexing with aminoclay. The experimental part consists of preparation of aminoclay complexes with selected bioactive agents at various concentrations. Verification of binding of bioactive agents to the aminoclay matrix was performed by Elemental Analysis (EA) and Fourier-transform infrared spectroscopy (FT-IR). For finding of the bound amount of bioactive substance were used the Ultra Performance Liquid Chromatography (UPLC®) and analysis of the cytotoxic properties of the formed complexes by used by the MTT assay. The main motivation of this study is to create new complexes with improved characteristics that would replace existing forms of substances used in pharmaceutical and biomedical applications.
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Synthesis of polymer precursors based on isosorbide
Šichnárek, Vojtěch ; Cigánek, Martin (referee) ; Figalla, Silvestr (advisor)
The theoretical part of the thesis deals with the description of reduced hexitols, methods of isosorbide production and its application purpose in polymer materials. It includes a description of the glucose dehydration process, the influence of reaction conditions and the catalysis of the isosorbide production, the polymerization of isosorbide monomers and their properties. The experimental part of the work is focused on the optimization of isosorbide’s laboratory synthesis‘ reaction conditions, the description of its chemical modification to polymerizable isosorbide derivatives and the physical-chemical characterization of the produced molecules. The synthesis of isosorbide is monitored by LC-MS, the product is subjected to various refining methods and a detailed structural characterization. GC was chosen for the monitoring of the preparation of isosorbide dimethacrylate and structural verification of the diester was confirmed by FT-IR, MS and NMR. The physical-chemical characterization of the prepared isosorbide dimethacrylate consists of the description of its rheological properties in mixture with a commercial methacrylated oil and the study of the reactivity of the monomers by DSC. The mixture of methacrylated oil and isosorbide dimethacrylate was used to prepare polymer thermosets which were analyzed by thermo-mechanical methods such as DMA, bending and TGA. The highest isosorbide conversion achieved value of 81.9 % after 5 hours, and it reached 94,1 % for isosorbide dimethacrylate after 8 hours. According to the results, isosorbide dimethacrylate possesses the abilty to reduce the shear viscosity of polymerizable precursors and to increase the glass transition temperature and crosslinking densities of polymer thermoset.
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Characterisation and chemical modification of halloysites
Vašutová, Vlasta ; Hradil, David (advisor) ; Pospíšil, Miroslav (referee)
Twelve halloysites from different sources in Slovakia, Turkey, China, New Zealand and U.S.A. have been characterized by combination of analytical methods together with the commercial sample of halloysite supplied by Sigma-Aldrich. The aim of this work was to select suitable candidates for to be used as carriers of porphyrine photoactive molecules. In nature, the formation of halloysite is related to the weathering of magmatic rocks or to the hydrothermal alteration of volcanic rocks, frequently in the contact with limestones. Halloysite belong to the kaolinite group, but, contrarily to kaolinite, it contains molecules of water in the interlayer space. It occurs in two forms: hydrated halloysite (10 ?) and dehydrated halloysite (7 ?).Dehydrated halloysites contain more admixtures than hydrated ones ? typically kaolinite, quartz, cristobalite, alunite, gibbsite and in one case also potassium mica. In samples containing both dehydrated halloysite and kaolinite their 001 diffractions overlap. Interaction with formamide was used in these cases to increase the interlayer space of halloysite and thus shift its basal diffraction to lower angles. The basal 001 diffraction of kaolinite after this treatment remains on 7 ?. Silver thiourea method (AgTU) was used to measure the cationic exchange capacity (CEC). Silver...
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Analytical methods for qualitative and quantitative determination of PHA in cyanobacteria
Černayová, Diana ; Samek, Ota (referee) ; Sedláček, Petr (advisor)
The diploma thesis is confused to verify the applicability of selected physicochemical and spectroscopic methods for characterization of cyanobacteria, with special emphasis on possibilities of qualitative and quantitative analysis of polyhydroxyalkanoates (specifically polyhydroxybutyrate (PHB)) accumulated in cyanobacterial cells. The sample basis of the work was formed by cultures of cyanobacterial strains of Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. The cultures were were cultivated in several ways to cover the widest possible range of physiological conditions and PHB contents, in particular using an autotrophic way of cultivation on shakers and multicultural culture method in a basic culture medium,and in media enriched with 2% salt (NaCl ) as well as mixotrophic culture media with different types of the carbon substrate. After few weeks of cultivation, cyanobacterial cultures were obtained and complexly analyzed by following techniques- cell suspensions were analyzed by flow cytometry and UV-VIS spectrometry (transmission and diffusion transmission mode), dry cell biomass was characterised by gas chromatography to obtain a exact amount of PHB, and then FT-IR spectrometry and thermogravimetric analysis. The work aimed to assess whether any of these methods can be a quick and affordable alternative to the determination of PHB content to the most commonly used method of gas chromatography, but also to assess what additional information about the physiological state of cyanobacterial cells can provide test methods. The highest correlation on PHB content was determined for the parameters determined by infrared spectroscopy, in which specific peaks from the characteristic wavelengths for polyhydroxybutyrate were important. Weak correlations on PHB content were achieved in thermogravimetric analysis and cytometry, using the hydrophobic fluorescent probe BODIPY 439/503, which bound to lipophilic parts of cells. In addition to the determination of PHB, it was possible to determine pigments present in cyanobacteria (such as chlorophyll, phycocyanin and carotenoids) by flow cytometry and UV-VIS diffusion transmission spectrometry. In the end, results from all used techniques were compared by PCA analysis to determine the similarity of all analyzed samples.
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Properties of aminoclay complexes and biologically active substances
Dušek, Jakub ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This paper builds on previous research of aminoclay complexes in undergraduate studies. Theoretical part deals with study of current problems of aminoclay complexes with bioactive substances and the choice of substances for complexing with aminoclay. The experimental part consists of preparation of aminoclay complexes with selected bioactive agents at various concentrations. Verification of binding of bioactive agents to the aminoclay matrix was performed by Elemental Analysis (EA) and Fourier-transform infrared spectroscopy (FT-IR). For finding of the bound amount of bioactive substance were used the Ultra Performance Liquid Chromatography (UPLC®) and analysis of the cytotoxic properties of the formed complexes by used by the MTT assay. The main motivation of this study is to create new complexes with improved characteristics that would replace existing forms of substances used in pharmaceutical and biomedical applications.
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